参桃软肝颗粒抑制肝癌的分子机制及调控RBPJ的探索  

Molecular Mechanism of Shentao Ruangan Granules in the Inhibition of Liver Cancer and its Exploration on Regulating RBPJ

作　　者：周瑞生 吴岱琳 高天琦[2] 黄丹 唐莹 周岱翰[1,3] ZHOU Rui-sheng;WU Dai-lin;GAO Tian-qi;HUANG Dan;TANG Ying;ZHOU Dai-han(The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;The First Clinical Medical School of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Institute of Oncology,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Technology Innovation Center,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学第一附属医院,广东广州510405 [2]广州中医药大学第一临床医学院,广东广州510405 [3]广州中医药大学肿瘤研究所,广东广州510405 [4]广州中医药大学科技创新中心,广东广州510405

出　　处：《中医肿瘤学杂志》2024年第1期49-56,共8页Journal of Oncology in Chinese Medicine

基　　金：广州中医药大学项目(编号:A1-2601-23-414-453Z73)。

摘　　要：目的肝细胞癌(hepatocellular carcinoma,HCC)是全球第四大常见癌症,严重威胁着国民健康。参桃软肝颗粒(STR)是国医大师周岱翰教授治疗肝癌的临床经验方,在临床实践中取得了良好的疗效,阐明其抑制肝癌生长和转移的机制是亟需解决的问题。方法Transwell侵袭迁移实验检测STR对HepG2细胞侵袭能力的影响,蛋白质印迹法(Western blot,WB)检测STR对Huh7细胞株RBPJ蛋白的表达影响。观察STR对Huh7荷瘤小鼠移植瘤生长的影响。RNA-seq鉴定STR处理的Huh7移植瘤组织中差异表达基因(differentially expressed genes,DEGs)。结果STR可显著抑制HepG2细胞的迁移和侵袭能力,STR可通过蛋白泛素化降解的途径减少RBPJ蛋白的表达;STR给药后,肝癌模型小鼠移植瘤体积以及瘤体重量均下降,肿瘤生长明显受到抑制。在对照组和STR给药组组间共发现2961个DEGs,其中上调和下调基因分别为1884和1077个。基因本体论(gene ontology,GO)、京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)和疾病本体论(disease ontology,DO)通路富集分析分别展示了STR可能作用的信号通路。结论Notch信号在肝癌中频繁发生异常表达及激活突变,转录因子RBPJ是Notch信号通路主要的下游靶基因,可以增强Notch信号通路中下游基因的表达。STR对肝细胞癌的抑制作用显著,其分子机制可能与抑制RBPJ的蛋白表达相关。Objective Hepatocellular carcinoma(HCC)is the fourth most common cancer in the world,posing a serious threat to national health.Shentao Ruangan Granules(STR)is a clinical experience prescription in the treatment of liver cancer by Professor Zhou Dai-han,a master of Chinese medicine.It has achieved good therapeutic effect in clinical practice,and illustrating its mechanism of inhibiting the growth and metastasis of liver cancer is an urgent issue needed to be solved.Methods The Transwell invasion and migration experiment was used to detect the effect of STR on the invasion ability of HepG2 cells,and Western blot(WB)was used to detect the effect on the expression of RBPJ protein in Huh7 cell line.The effect of STR on the growth of transplanted tumor in Huh7 tumor bearing mice was observed.Differentially expressed genes(DEGs)of Huh7 transplanted tumor tissue treated with STR were identified by RNA-seq.Results STR could significantly inhibited the migration and invasion ability of HepG2 cells and reduce the expression of RBPJ protein via protein ubiquitination degradation.After STR treatment,the volume and weight of transplanted tumors in liver cancer model mice decreased,and the growth of tumors was significantly inhibited.A total of 2961 DEGs were found between the control group and the STR treatment group,with 1884 up-regulated genes and 1077 down-regulated genes,respectively.Gene ontology(GO),Kyoto encyclopedia of genes and genomes(KEGG),and disease ontology(DO)pathway enrichment analysis showed the possible signaling pathways of STR respectively.Conclusion Notch signaling experiences abnormal expression and activation mutations in liver cancer frequently.The transcription factor RBPJ is the main downstream target gene of Notch signaling pathway,which can enhance the expression of downstream genes in the Notch signaling pathway.STR has a significant inhibitory effect on hepatocellular carcinoma,and its molecular mechanism may be related to inhibit the expression of RBPJ protein.

关 键 词：参桃软肝颗粒 肝细胞癌 RBPJ RNA-seq测序 分子机制 

分 类 号：R735.7[医药卫生—肿瘤] R730.52[医药卫生—临床医学] R273